Evidence for copurification of micronuclei in sucrose density gradient-enriched plasma membranes from cell lines
| Autor: | Xuejun Sun, Jennifer Dufour, Liang Li, Cheryl Santos, Lidan Tao, Carol E. Cass, John R. Mackey, Vijaya L. Damaraju, Nan Li, Sambasivarao Damaraju, David S. Wishart, Nan Zhang |
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| Rok vydání: | 2010 |
| Předmět: |
Biophysics
Biology Biochemistry Copurification Cell membrane Mice 03 medical and health sciences 0302 clinical medicine Cell Line Tumor Centrifugation Density Gradient medicine Animals Humans Centrifugation Nuclear protein Molecular Biology Micronuclei Chromosome-Defective Cellular compartment 030304 developmental biology 0303 health sciences Cell Membrane Nuclear Proteins DNA Neoplasm Cell Biology Alkaline Phosphatase Molecular biology Membrane medicine.anatomical_structure Membrane protein 030220 oncology & carcinogenesis Cell fractionation Biomarkers Subcellular Fractions |
| Zdroj: | Analytical Biochemistry. 396:69-75 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2009.08.019 |
| Popis: | Sucrose density gradient-enriched membrane preparations and membrane fraction enrichment through affinity purification techniques are commonly used in proteomic analysis. However, published proteomic profiles characterized by the above methods show the presence of nuclear proteins in addition to membrane proteins. While shuttling of nuclear proteins across cellular compartments and their transient residency at membrane interfaces could explain some of these observations, the presence of nuclear proteins in proteomic profiles generated with crude and enriched membranes could be the result of nonspecific contamination of nuclear debris during cell fractionation procedures. We hypothesized that micronuclei arising from the genomic instability inherent to cancer cells may copurify with plasma membrane fractions on sucrose gradients. Using sucrose gradient-enriched plasma membranes from breast cancer cell lines derived from the MCF-7 cell line, we provide experimental evidence to indicate that micronuclei are present in fresh preparations of plasma membranes. The origin of these micronuclei was traced to budding of nuclei in intact cells. Furthermore, mass spectrometric analysis confirmed the presence of nuclear proteins as well as membrane and associated signaling proteins in sucrose gradient-enriched preparations. |
| Databáze: | OpenAIRE |
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